Experimental autoimmune uveoretinitis (EAU) is a T-cell-mediated autoimmune disease that serves as a model of human intraocular inflammatory disease (uveitis). It is initiated in susceptible animals by immunization with retinal antigens, such as interphotoreceptor retinoid-binding protein (IRBP) and S-antigen (SAg). Previous studies of T-cell receptor (TCR) usage by uveitogenic T cells suggested a possible connection between pathogenicity of T lymphocytes and usage of V~8 family genes. Here, we have analyzed the T-cell repertoire at the autoimmune site by examining V~ gene expression in the retinas of animals with SAg- or IRBP-induced uveitis. Our data show a remarkable bias for the use of V~8.2 TCR among V~8-positive T cells in the retinas of animals with SAg-EAU, while in IRBP-EAU both V~8.2-positive and V~8.3-positive T cells were found. Similar results were obtained by DNA sequence analysis of 75 V~8 cDNA clones from uveitogenic T-cell lines. Twenty clones isolated from SAg-specific T-cell lines were all V~8.2 TCRs, whereas, among 55 V~8 cDNAs from IRBP-specific lines, 22% were V~8.2; 33%, V~8.3; and 44%, V~8.2 isoforms not previously reported. V~8.1-positive T cells were absent from the V~8 repertoires of the retina or any of the T- cell lines. In addition, there was marked heterogeneity of the CDR3 sequences utilized by the V~8-positive uveitogenic T cells, which contrasts with the reported near homogeneity of encephalitogenic T cells involved in experimental allergic encephalomyelitis (EAE). Taken together, our data show that although there is a bias toward usage of V~8-positive cells, the T-cell response in EAU is not oligoclonal. Moreover, distinct V~8 subfamilies were differentially activated by the autoantigens SAg and IRBP. In view of these findings, we believe that anti-V~ therapy might have only limited usefulness in human uveitis because several autoantigens and an unknown number of immunopathogenic epitopes may be involved in the etiology of this diverse group of diseases.